In vivo imaging of retrovirus infection reveals a role for Siglec-1/CD169 in multiple routes of transmission

Haugh, Kelsey A.; Ladinsky, Mark S.; Ullah, Irfan; Stone, Helen; Pi, Ruoxi; Gilardet, Alexandre; Grunst, Michael W; Kumar, Priti; Björkman, Pamela J.; Mothes, Walther; Uchil, Pradeep D.

doi:10.7554/elife.64179

Cited by 9 publications

(9 citation statements)

References 57 publications

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“…Thus, our work shows that the gammaretrovirus MLV can infect non-dividing cells like DCs, thought to be the initial in vivo targets of MLV [12, 13, 43]. Interestingly, previous studies have suggested that gammaretroviruses can infect other quiescent cell types, such as neurons [44].…”

Section: Discussionmentioning

confidence: 79%

Murine leukemia virus can infect non-dividing cells

Salas-Briceno,

Zhao,

Ross

2023

Preprint

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Retroviral reverse transcription starts within the capsid and uncoating and reverse transcription are mutually dependent. There is still debate regarding the timing and cellular location of HIV’s uncoating and reverse transcription and whether it occurs solely in the cytoplasm, nucleus or both. HIV can infect non-dividing cells because there is active transport of the preintegration complex (PIC) across the nuclear membrane, but Murine Leukemia Virus (MLV) is thought to depend on cell division for replication and whether MLV uncoating and reverse transcription is solely cytoplasmic has not been studied. Here, we used NIH3T3 and primary mouse dendritic cells to determine where the different stages of reverse transcription occur and whether cell division is needed for nuclear entry. Our data strongly suggest that in both NIH3T3 cells and dendritic cells (DCs), the initial step of reverse transcription occurs in the cytoplasm. However, we detected MLV RNA/DNA hybrid intermediates in the nucleus of dividing NIH3T3 cells and non-dividing DCs, suggesting that reverse transcription can continue after nuclear entry. We also found that the MLV PIC requires cell division to enter the nucleus of NIH3T3 cells. In contrast, we show that MLV can infect non-dividing primary DCs, although integration of MLV DNA in DCs still required the viral p12 protein. Knockdown of several nuclear pore proteins dramatically reduced the appearance of integrated MLV DNA in DCs but not NIH3T3 cells. Additionally, MLV capsid associates with the nuclear pore proteins NUP358 and NUP62 during infection. These findings suggest that simple retroviruses, like HIV, gain nuclear entry by traversing the nuclear pore complex in non-mitotic cells.

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Section: Discussionmentioning

confidence: 79%

Murine leukemia virus can infect non-dividing cells

Salas-Briceno,

Zhao,

Ross

2023

Preprint

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show abstract

“…Thus, our work shows that the gammaretrovirus MLV can infect non-dividing cells like DCs, thought to be the initial in vivo targets of MLV [ 12 , 13 , 44 ]. Interestingly, previous studies have suggested that gammaretroviruses can infect other quiescent cell types, such as neurons [ 45 ].…”

Section: Discussionmentioning

confidence: 99%

Murine leukemia virus infection of non-dividing dendritic cells is dependent on nucleoporins

Salas-Briceno,

Zhao,

Ross

2024

PLoS Pathog

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Retroviral reverse transcription starts within the capsid and uncoating and reverse transcription are mutually dependent. There is still debate regarding the timing and cellular location of HIV’s uncoating and reverse transcription and whether it occurs solely in the cytoplasm, nucleus or both. HIV can infect non-dividing cells because there is active transport of the preintegration complex (PIC) across the nuclear membrane, but Murine Leukemia Virus (MLV) is thought to depend on cell division for replication and whether MLV uncoating and reverse transcription is solely cytoplasmic has not been studied. Here, we used NIH3T3 and primary mouse dendritic cells to determine where the different stages of reverse transcription occur and whether cell division is needed for nuclear entry. Our data strongly suggest that in both NIH3T3 cells and dendritic cells (DCs), the initial step of reverse transcription occurs in the cytoplasm. However, we detected MLV RNA/DNA hybrid intermediates in the nucleus of dividing NIH3T3 cells and non-dividing DCs, suggesting that reverse transcription can continue after nuclear entry. We also confirmed that the MLV PIC requires cell division to enter the nucleus of NIH3T3 cells. In contrast, we show that MLV can infect non-dividing primary DCs, although integration of MLV DNA in DCs still required the viral p12 protein. Knockdown of several nuclear pore proteins dramatically reduced the appearance of integrated MLV DNA in DCs but not NIH3T3 cells. Additionally, MLV capsid associated with the nuclear pore proteins NUP358 and NUP62 during infection. These findings suggest that simple retroviruses, like the complex retrovirus HIV, gain nuclear entry by traversing the nuclear pore complex in non-mitotic cells.

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“…Subsequently, we adoptively transfected DiR-labeled spleen-derived iNKT cells with specific phenotypes and functions into normal DBA/1 mice and tracked the migration, distribution, and metabolism of iNKT cells in mice using a small animal imaging technique. Previous studies have reported that tail vein injection results in fluorescence in organs such as the liver and spleen within 30 seconds [ 26 ]. We observed that strong fluorescence can be detected in the lungs immediately after cell infusion, and the fluorescence intensity was the strongest at 10 min and then gradually weakened.…”

Section: Discussionmentioning

confidence: 99%

Migration, Distribution, and Safety Evaluation of Specific Phenotypic and Functional Mouse Spleen-Derived Invariant Natural Killer T2 Cells after Adoptive Infusion

Chen

Teng

et al. 2021

Mediators of Inflammation

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Herein, the migration distribution and safety of specific phenotypic and functionally identified spleen-derived invariant natural killer T2 (iNKT2) cells after adoptive infusion in mice were studied. The proliferation and differentiation of iNKT cells were induced by intraperitoneal injection of α-galactosylceramide (α-GalCer) in vivo. Mouse spleens were isolated in a sterile environment. iNKT cells were isolated by magnetic-activated cell sorting columns (MS columns). Cytometric bead array (CBA) assay was used to detect cytokine secretion in the supernatant stimulated by iNKT cells. The basic life status of the mice was observed, and systematic quantitative scoring was conducted after injecting spleen-derived iNKT cells through the tail vein. An in vivo imaging system was used to trace the migration and distribution of iNKT cells in DBA mice. The percentage of the iNKT2 subgroup was the highest in 3 days after intraperitoneal injection of α-GalCer, and iNKT2 subsets accounted for more than 92% after separation and purification by magnetic-activated cell sorting (MACS). Anti-inflammatory cytokine IL-4 was mainly found in the supernatant of cell cultures. The adoptive infusion of iNKT cells into healthy mice resulted in no significant change in the basic life status of mice compared with the noninjected group. iNKT cells were detected in the lung, spleen, and liver, but no fluorescence was detected in lymph nodes and thymus. After dissecting the mice, it was found that there were no significant abnormalities in the relevant immune organs, brain, heart, kidney, lung, and other organs. Intraperitoneal injection of α-GalCer results in a large number of iNKT2 cells, mainly secreting anti-inflammatory cytokine IL-4, from the spleen of mice. After adoptive infusion, the iNKT2 cells mainly settled in the liver and spleen of mice with a satisfactory safety profile.

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In vivo imaging of retrovirus infection reveals a role for Siglec-1/CD169 in multiple routes of transmission

Cited by 9 publications

References 57 publications

Murine leukemia virus can infect non-dividing cells

Murine leukemia virus can infect non-dividing cells

Murine leukemia virus infection of non-dividing dendritic cells is dependent on nucleoporins

Migration, Distribution, and Safety Evaluation of Specific Phenotypic and Functional Mouse Spleen-Derived Invariant Natural Killer T2 Cells after Adoptive Infusion

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